Article

1H, 13C, and 15N resonance assignment of the central domain of hRSV transcription antitermination factor M2-1.

Unité de Virologie et Immunologie Moléculaires, INRA UR0892, 78350, Jouy-en-Josas, France.
Biomolecular NMR Assignments (impact factor: 0.72). 04/2011; 5(2):237-9. DOI:10.1007/s12104-011-9308-3 pp.237-9
Source: PubMed

ABSTRACT M2-1 is an essential co-factor of the respiratory syncytial virus, an important respiratory pathogen in infants and calves. It acts as a transcription antitermination factor which enhances the processivity of the polymerase. Within the polymerase complex, M2-1 interacts with a second co-factor, the phosphoprotein P. It has been shown previously that P and RNA bind to M2-1 in a competitive manner in vitro and that these properties are related to a central domain located between residues Glu59 and Lys177. Here we report the almost complete (1)H, (13)C and (15)N assignment of a fragment of M2-1 corresponding to this region, for further structure determination and interaction studies.

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    Article: Structure and functional analysis of the RNA- and viral phosphoprotein-binding domain of respiratory syncytial virus M2-1 protein.
    Marie-Lise Blondot, Virginie Dubosclard, Jenna Fix, Safa Lassoued, Magali Aumont-Nicaise, François Bontems, Jean-François Eléouët, Christina Sizun
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    ABSTRACT: Respiratory syncytial virus (RSV) protein M2-1 functions as an essential transcriptional cofactor of the viral RNA-dependent RNA polymerase (RdRp) complex by increasing polymerase processivity. M2-1 is a modular RNA binding protein that also interacts with the viral phosphoprotein P, another component of the RdRp complex. These binding properties are related to the core region of M2-1 encompassing residues S58 to K177. Here we report the NMR structure of the RSV M2-1(58-177) core domain, which is structurally homologous to the C-terminal domain of Ebola virus VP30, a transcription co-factor sharing functional similarity with M2-1. The partial overlap of RNA and P interaction surfaces on M2-1(58-177), as determined by NMR, rationalizes the previously observed competitive behavior of RNA versus P. Using site-directed mutagenesis, we identified eight residues located on these surfaces that are critical for an efficient transcription activity of the RdRp complex. Single mutations of these residues disrupted specifically either P or RNA binding to M2-1 in vitro. M2-1 recruitment to cytoplasmic inclusion bodies, which are regarded as sites of viral RNA synthesis, was impaired by mutations affecting only binding to P, but not to RNA, suggesting that M2-1 is associated to the holonucleocapsid by interacting with P. These results reveal that RNA and P binding to M2-1 can be uncoupled and that both are critical for the transcriptional antitermination function of M2-1.
    PLoS Pathogens 05/2012; 8(5):e1002734. · 9.13 Impact Factor
Data provided are for informational purposes only. Although carefully collected, accuracy cannot be guaranteed. The impact factor represents a rough estimation of the journal's impact factor and does not reflect the actual current impact factor. Publisher conditions are provided by RoMEO. Differing provisions from the publisher's actual policy or licence agreement may be applicable.

Keywords

15)N assignment
 
central domain
 
essential co-factor
 
infants
 
interaction studies
 
M2-1 corresponding
 
M2-1 interacts
 
polymerase
 
polymerase complex
 
processivity
 
RNA bind
 
second co-factor
 
transcription antitermination factor
 
vitro